eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2025-12-30 22 4 1580 1586 10.13005/bbra/3461 56758 article Shailja Jitendrabhai Vyas 1 Anjali Kulkarni 2 Department of Genetics, Ashok and Rita Patel Institute of Integrated Study and Research in Biotechnology and Allied Science (ARIBAS), Vallabh Vidyanagar, Anand, Gujarat. Department of Biotechnology, Genetics and Bioinformatics, Natubhai V. Patel College of Pure and Applied Sciences (NVPAS), Vallabh Vidhyanagar, Anand, Gujarat. Bovine milk and dairy products are vital components of traditional diets globally, with milk proteins comprising approximately 80% casein and 20% whey. Among the caseins, β-casein is a major hydrophobic protein with two common genetic variants, A1 and A2. The difference between these variants lies in a single amino acid substitution at position 67, where histidine (CCT) in A1 is replaced by proline (CAT) in A2. This study aimed to investigate the β-casein gene polymorphism in 200 Gir cattle from Shri Krishna Farm, Savarkundla  using the PCR-RFLP technique. Genomic DNA was extracted using the Phenol-Chloroform-Isoamyl Alcohol method, and its quality and quantity were assessed through agarose gel electrophoresis and Nanodrop spectrophotometry. PCR amplification yielded a 121 bp fragment, which was then subjected to restriction digestion using the DdeI enzyme. The analysis revealed two genotypes—A1A2 and A2A2—with respective genotypic frequencies of 0.125 and 0.875, while the A1A1 genotype was not observed. The allele frequencies were 0.06 for A1 and 0.94 for A2, confirming the predominance of the A2 allele in Gir cattle and suggesting their potential for producing A2 milk, which is considered healthier. https://www.biotech-asia.org/vol22no4/study-of-beta-casein-gene-polymorphism-in-gir-cattle-from-gujarat-state/ Allele Frequency; Beta-Casein; Genotype; Milk; PCR-RFLP